Santhosh Gatreddi

18 Questions 53 Answers 0 Followers

Questions related from Santhosh Gatreddi

How to determine the amount of acid required to stop the enzyme reaction?

I have to stop my enzyme reaction pH(8.5) by perchloric acid and further neutralized by KHCO3. How to determine the amount of acid that is required to stop the reaction and is there any...

06 June 2017 855 5 View

How to calculate the slope of a line that resembles the exponential decay?

Iam measuring the absorption of substrate in a enzymatic reaction to find out the kinetic parameters Km and Vmax. Plot of the curve (absorbance vs time) obtained in a exponential decay manner. To...

12 December 2016 8,895 3 View

How can I dissolve ZnCl2 in water?

Iam trying to prepare 0.1M ZnCl2 solution but it is forming white precipitate. What are factors that influence its solubility in water or what might be the probable solvent to make it soluble.

05 May 2015 4,909 8 View

Is it possible to crystallize proteins which precipitate immediately after purification?

I have read in one of the article that they have crystallized protein by screening the protein solution which was partially aggregated before even setting and the drop and finally they got good...

04 April 2015 5,934 8 View

How might one overcome protein precipitation during elution ?

I have purified a protein of 36kDa with Ni-NTA affinity chrmotography and initially protein was eluted with 100-500mM imidazole. When elution was started with 300mM imidazole and so on protein...

11 November 2014 9,019 13 View

How can I eliminate the interference of sarcosine in the binding of protein to ni-nta?

I have solubilized my protein with 0.3% sarcosine and purified by using Ni-NTA,during purification most protein is going into flow through. I have diluted my sonicated sample to 0.1% sarcosine...

05 May 2014 8,238 5 View

Anyone familiar with the fate of expressed protein in E.coli?

If I have expressed a functional protein in e.coli,how the activity of the protein in bacterial cytoplasm will effect its survival? If it is a metabolic enzyme and by chance it is involved in...

05 May 2014 9,732 2 View

What is the minimum percentage of sarkosyl that is allowed for a lysis buffer when extracting native proteins from inclusion bodies?

I have sonicated my bacterial cell pellet with 0.5% sarkosyl and have a sufficient amount in soluble fraction. In another condition, I have used 50mM each of L-Arginine and glycyl glycine in...

04 April 2014 3,008 3 View

I have purified my protein coupled to gst tag and i got several non specific bands during purification, how to reduce non-specificity ?

My protein has 59 kDa and I got gst along with it during purification. I am confused whether my protein is degrading or my induced colony has both native and/or vector with insert.

12 December 2013 8,570 1 View

I have a protein concentrator of 30kDa and my protein of interest has 35kDa, is it possible to purify my protein with this?

If not, which is the better concentrator that is suitable for the exclusion of non-specific bands (at 20kDa)?

09 September 2013 8,809 13 View

Could someone suggest some methods for the purification of proteins from inclusion bodies?

As most of the proteins are going into the insoluble fraction as an aggregate and a very low amount in soluble fraction...

09 September 2013 661 5 View

Is it possible to use E.coli BL21(DE3) as a host for the expression of my target protein cloned in pQE 30 vector?

I have read some articles, in which they have used BL21 and in other some cases it is M15 cells. So, which is the better host strain for expression and why ?

09 September 2013 5,575 9 View

Is there any chance for the pelleting of soluble protein during centrifugation (due to centrifugal force)?

I have taken my sonicated sample and kept it for centrifugation (10000rpm for 15min) to separate the pellet from supernatant. After that, I centrifuged my supernatant @15000rpm for 30 min and...

05 May 2013 6,249 36 View

Why bacterial cells were not sonicating properly after a long duration of sonication?

I have used BL21 DE3 for expression. After overnight induction at 16°C, I have harvested the cells and sonicated at 40% amplitude for 30sec on/off for several cycles, but still my cells not lysing...

05 May 2013 9,190 16 View

Which buffer is good to do protein purification by Ni-NTA?

Which buffer is good for buffering capacity and interaction with protein?

05 May 2013 7,753 1 View

How to assay the activity of the ser/thr protein phosphatase?

If anybody has the protocol, please forward it to me.

05 May 2013 3,908 0 View

How to overcome the elution of non-specific bands in protein purification by Ni-NTA (with His-tag)?

I have eluted my protein with 200mM imidazole and got some non specific bands in the elution but concentration is good. Elution with 300mM imidazole leads to the reduction of non-specific protein...

05 May 2013 10,408 31 View

How does one choose the pH of the buffer during purification of recombinant proteins based on its isoelectric point?

How does the isoelectric pH of a protein effect its solubility during its purification from bacteria?

05 May 2013 4,292 2 View